González L M, Daniel-Mwambete K, Montero E, Rosenzvit M C, McManus D P, Gárate T, Cuesta-Bandera C
Instituto de Salud Carlos III, Centro Nacional de Microbiología, 28220 Majadahonda, Madrid, Spain.
Exp Parasitol. 2002 Sep;102(1):46-56. doi: 10.1016/s0014-4894(02)00146-7.
We have designed two polymerase chain reaction (PCR) primer sets (PEg9F1-PEg9R1 and PEg16F1-PEg16R1) and two PCR protocols (Eg9-PCR and Eg16-PCR) for discrimination of Echinococcus granulosus genotypes. The oligonucleotide sequences originate from two E. granulosus DNA multiplex-PCR amplification fragments, previously reported, that allows species-specific discrimination between Taenia saginata, Taenia solium, and E. granulosus. The Eg9-PCR, Eg16-PCR, and Eg9-PCR linked restriction fragment length polymorphism (RFLP) analysis was used to characterize 53 E. granulosus isolates from the central region of Spain, highly endemic for echinococcosis. The analysis resulted in: (i) the discrimination of E. granulosus from Echinococcus multilocularis; (ii) the characterisation and discrimination of discrete E. granulosus strains from Spain; and (iii) the identification of two distinct genotypes within E. granulosus Spanish pig isolates. To further characterize the genetic variants in pigs, fragments of the NADH dehydrogenase I (ND1) and the cytochrome c oxidase subunit I (CO1) genes were amplified from parasite DNA and sequenced. The results again revealed the presence of two distinct genotypes: the G1 (sheep-dog strain) and G7 (pig-dog strain) genotypes. This observation could have important consequences for human health in Spain. Furthermore, the Eg9-PCR, Eg16-PCR, and Eg9-PCR-RFLP protocols can be used as additional methods to discriminate various E. granulosus genotypes.
我们设计了两套聚合酶链反应（PCR）引物组（PEg9F1-PEg9R1和PEg16F1-PEg16R1）以及两种PCR方案（Eg9-PCR和Eg16-PCR），用于鉴别细粒棘球绦虫的基因型。这些寡核苷酸序列源自先前报道的两个细粒棘球绦虫DNA多重PCR扩增片段，可实现牛带绦虫、猪带绦虫和细粒棘球绦虫之间的种特异性鉴别。使用Eg9-PCR、Eg16-PCR以及Eg9-PCR连锁限制性片段长度多态性（RFLP）分析对来自西班牙中部地区的53株细粒棘球绦虫分离株进行特征分析，该地区是棘球蚴病的高度流行区。分析结果如下：（i）鉴别细粒棘球绦虫和多房棘球绦虫；（ii）对来自西班牙的不同细粒棘球绦虫菌株进行特征分析和鉴别；（iii）在西班牙猪分离株的细粒棘球绦虫中鉴定出两种不同的基因型。为了进一步表征猪体内的遗传变异，从寄生虫DNA中扩增出烟酰胺腺嘌呤二核苷酸脱氢酶I（ND1）和细胞色素c氧化酶亚基I（CO1）基因的片段并进行测序。结果再次揭示存在两种不同的基因型：G1（绵羊-犬株）和G7（猪-犬株）基因型。这一观察结果可能对西班牙的人类健康产生重要影响。此外，Eg9-PCR、Eg16-PCR和Eg9-PCR-RFLP方案可作为鉴别各种细粒棘球绦虫基因型的补充方法。
